Combined RT-qPCR of mRNA and microRNA targets within one fluidigm integrated fluidic circuit

Don A. Baldwin, Annamarie D. Horan, Patrick J. Hesketh, Samir Mehta

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The ability to profile expression levels of a large number of mRNAs and microRNAs (miRNAs) within the same sample, using a single assay method, would facilitate investigations of miRNA effects on mRNA abundance and streamline biomarker screening across multiple RNA classes. A protocol is described for reverse transcription of long RNA and miRNA targets, followed by preassay amplification of the pooled cDNAs and quantitative PCR (qPCR) detection for a mixed panel of candidate RNA biomarkers. The method provides flexibility for designing custom target panels, is robust over a range of input RNA amounts, and demonstrated a high assay success rate.

Original languageEnglish
Pages (from-to)75-83
Number of pages9
JournalJournal of Biomolecular Techniques
Volume27
Issue number2
DOIs
StatePublished - Jul 2016

Keywords

  • Adult
  • Biomarkers/blood
  • Case-Control Studies
  • Gene Expression Profiling/instrumentation
  • Humans
  • MicroRNAs/blood
  • Microfluidic Analytical Techniques
  • RNA, Messenger/blood
  • Reverse Transcriptase Polymerase Chain Reaction

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